Invitrogen Qubit 4 User Manual
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Invitrogen Qubit 4 User Manual

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Qubit
4 Fluorometer
™
Catalog Number Q33226
Publication Number MAN0017209
Revision A.0
For Research Use Only. Not for use in diagnostic procedures.

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Summary of Contents for Invitrogen Qubit 4

  • Page 1 Qubit 4 Fluorometer ™ Catalog Number Q33226 Publication Number MAN0017209 Revision A.0 For Research Use Only. Not for use in diagnostic procedures.
  • Page 2 Information in this document is subject to change without notice. DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.

  • Page 3: Table Of Contents

    Contents About this guide ......................3 1. Product information ....................5 Product contents .............................. 5 Product description ............................6 Instrument exterior components ........................7 Product specifications ............................. 8 2. Getting started ......................9 Set up the Qubit ™ 4 Fluorometer ........................9 Optional: Configure instrument settings .....................
  • Page 4 Appendix A: Troubleshooting..................55 Troubleshooting ............................. 55 Critical Qubit ™ Assay considerations ......................57 Appendix B: Ordering information ................59 Qubit 4 Fluorometer and accessories ....................... 59 ™ Appendix C: Safety ...................... 60 Symbols on instruments ..........................60 Safety labels on instruments ........................61 General instrument safety ..........................

  • Page 5: About This Guide

    About this guide Overview This user guide describes how to operate the Qubit ™ 4 Fluorometer. User The guides listed below are available with the Qubit 4 Fluorometer. ™ documentation Guide Pub. No. Qubit ™ 4 Fluorometer User Guide MAN0017209 Qubit ™...
  • Page 6 Safety alert words Four safety alert words appear in Thermo Fisher Scientific user documentation at points in the document where you need to be aware of relevant hazards. Each alert word—IMPORTANT, CAUTION, WARNING, DANGER—implies a particular level of observation or action, as defined below: IMPORTANT! –...

  • Page 7: Product Information

    1. Product information Product contents The Qubit 4 Fluorometer is shipped with the components listed below: ™ Component Quantity Qubit 4 Fluorometer 1 each ™ Universal Power Cord with 4 adaptor plugs 1 each (for U.S./E.U./U.K./Australia) USB drive (includes the user guide) 1 each Qubit ™...

  • Page 8: Product Description

    Product description Qubit The Qubit ™ 4 Fluorometer is a benchtop fluorometer that can be used for the ™ quantitation of DNA, RNA, microRNA, and protein, as well as for the measurement Fluorometer of RNA integrity and quality using the highly sensitive and accurate fluorescence- assays.

  • Page 9: Instrument Exterior Components

    Instrument exterior components Top view Back view Touchscreen is the user interface containing the controls for all the functions needed and displays data from the assays. Sample chamber is used to load the assay tube containing the sample into the fluorometer for analysis.

  • Page 10: Product Specifications

    Product specifications Physical Instrument type: Benchtop fluorometer characteristics Instrument dimensions: 5.4 in (w) × 10 in (l) × 2.2 in (h) (13.6 cm × 25 cm × 5.5 cm); rectangular shape Weight: 26.2 oz (743 g) Operating power: 100–240 VAC, 1.0 A/1.2 A Frequency: 50/60 Hz Electrical input:...

  • Page 11: Getting Started

    2. Getting started Set up the Qubit 4 Fluorometer ™ Set up the The Qubit 4 Fluorometer is a stand-alone instrument that does not require ™ connection to a computer. instrument After unpacking the instrument, place the instrument on a flat, level, dry surface.
  • Page 12 About Instrument The About Instrument screen displays information about your Qubit ™ 4 Fluorometer, including the currently installed software version. To access the About Instrument screen screen: On the Home screen, press Settings. On the Settings screen, press About Instrument to display the About Instrument screen.

  • Page 13: Optional: Configure Instrument Settings

    Optional : Configure instrument settings Introduction You can configure the following instrument settings for the Qubit ™ 4 Fluorometer: • Sleep mode (page 12) • Brightness (page 12) • Date/Time (page 13) • Reset instrument (page 14) • Language (page 15) Access the On the Home screen, press Settings.
  • Page 14 Adjust the sleep The Qubit ™ 4 Fluorometer has a sleep mode (i.e., automatic standby) that is triggered by inactivity. The system default is 10 minutes of inactivity before the mode instrument goes into sleep mode. To adjust this time: On the Instrument settings screen (page 11), press Sleep mode to display the Sleep mode screen.
  • Page 15 Adjust the date and The date and time are preset when you receive the instrument. To adjust the date and time: time On the Instrument settings screen (page 11), press Date/Time.  Select a date format, select a time format, then press Next. Enter the date and time: Press a date field (day, month, or year), then use the number pad to select a value.
  • Page 16 Reset function The Reset function returns the Qubit ™ 4 Fluorometer to its default factory settings, erasing all saved data, user–defined instrument settings, and custom assays. IMPORTANT! The reset function is not reversible. To return the instrument to its default factory settings: On the Instrument settings screen (page 11), press Reset instrument to display the Reset Qubit 4 screen.
  • Page 17 Change the display You can change the language that the Qubit ™ 4 Fluorometer displays to English (default), French, German, Italian, Spanish, simplified Chinese, and Japanese. language On the Instrument settings screen (page 11), press Language to display the Language screen. ...

  • Page 18: Guidelines For Using The Qubit ™ 4 Fluorometer

    Guidelines for using the Qubit 4 Fluorometer ™ Recommendations To obtain the best results, follow the recommendations below. For more information, see “Critical Qubit ™ Assay considerations”, page 57. • Do not operate the instrument in direct sunlight. • Wear gloves during sample handling. •...

  • Page 19: Perform Assays

    3. Perform assays Overview This section provides instructions for using the Reagent Calculator, running new calibration standards or for using the last calibration, and for reading the samples using a Qubit ™ assay kit. Materials needed • A Qubit ™ assay kit appropriate for quantifying your samples (see page 59 for available Qubit assay kits and ordering information)

  • Page 20: Optional: Use Reagent Calculator To Prepare Qubit ™ Working Solution

    Optional : Use Reagent Calculator to prepare Qubit Working Solution ™ Introduction Use the on-board Reagent Calculator to quickly determine the correct amount of Qubit ™ dye and buffer required to make the appropriate amount of Qubit ™ Working Solution required to prepare your samples and standards. Use the Reagent On the Home screen, press Reagent Calculator.
  • Page 21 Press Enter to calculate the amount of Qubit ™ dye and buffer required to prepare the Qubit ™ Working Solution with these inputs. Note: You can change the total number of tubes that you plan to run or the overage selection on this screen. Press Done to return to the Home screen to run your Qubit assay.

  • Page 22: Calibrate Assays On The Qubit ™ 4 Fluorometer

    Calibrate assays on the Qubit 4 Fluorometer ™ Introduction For each assay, you have the choice to run new standards for calibrating the assay on the Qubit ™ 4 Fluorometer or to use the values from the previous calibration (for more information, see “Qubit™...
  • Page 23 If you have already performed a calibration for the selected assay, the software prompts you to choose between reading new standards and running samples using the previous calibration. Press Read standards. Note: To apply the previous calibration to your sample readings, press Run samples.
  • Page 24 Calibration results • For Qubit quantitation assays: If the calibration is successful, the software ™ displays the Read standard screen with the Fluorescence vs. Concentration graph. In the Fluorescence vs. Concentration graph, the standard data points are connected by a line and open circles represent correct standards. •...
  • Page 25 Calibration error If you receive the “Calibration error” message, you can re-run the standards: In the Error screen, press OK. Review the Read standard screen (see Note on page 22). If you wish to re-run the standards, or run new standards, press Read standards, then repeat the calibration procedure (beginning on page 18).

  • Page 26: Read Samples

    Read samples Before you begin • Calibrate the Qubit 4 Fluorometer as described on page 18. (Run the ™ appropriate standards or accept the values from the previous calibration.) • Prepare the samples. Refer to the instructions provided with the assay. Note: Incubate the samples for the appropriate amount of time after mixing them with the working solution (2 minutes for the Qubit ™...
  • Page 27 Insert a sample tube into the sample chamber, close the lid, and then press Read tube. The reading takes approximately 3 seconds for the quantitation assays and approximately 5 seconds for the quality assays.  The software displays the results. See “Results screen” on page 26. Results screen for quantitation Results screen for quality assays...

  • Page 28: Results

    Results Results screen – The Results screen displays the results of the sample run. Sample • If the results are within the assay’s range, the concentration values are displayed. quantitation The top value (in large font) is the concentration of the original sample. The bottom value is the dilution concentration (the concentration of the sample in the tube inserted into the Qubit 4 fluorometer).
  • Page 29 • To view the Fluorescence vs. Concentration graph, swipe left or press the right arrow. In the graph: ­ Open circles represent correct standards. ­ The large gray circle represents the most recent sample. ­ Blue circles represent samples that fall within the assay’s core range. ­...
  • Page 30 Results screen – The Results screen displays the results of the sample run. Sample quality • If the results are within the assay’s range, the sample quality values are displayed. The result presented in the circle (i.e., RNA IQ#) is the total value for the RNA sample integrity and quality, which is the calculated percentage of large and small RNA in the sample.

  • Page 31: Fluorometer Mode

    Fluorometer mode Introduction You can use the Qubit ™ 4 Fluorometer as a mini-fluorometer by selecting the Fluorometer mode. The Fluorometer mode allows you to choose the excitation light source (blue LED or red LED) while reading fluorescence in both the green and far-red emission channels (for the blue LED) or in the far-red emission channel only (for the red LED).
  • Page 32 Insert the sample, then press Read tube. For Blue excitation, the Qubit ™ 4 Fluorometer displays both the green emission and the far red emission of your sample. For Red excitation, the Qubit ™ 4 Fluorometer displays only the far red emission of your sample. Blue (470 nm) excitation Red (635 nm) excitation Read multiple...

  • Page 33: Ion Sphere ™ Assay

    Ion Sphere Assay ™ Ion Sphere Assay The Ion Sphere ™ assay on the Qubit ™ 4 Fluorometer enables quick and easy ™ assessment of Ion Sphere ™ Particles (ISPs) prior to performing a sequencing run on the Ion Personal Genome Machine ™...

  • Page 34: Manage Data

    4. Manage data Introduction The Qubit 4 Fluorometer can save data for up to 1000 samples. ™ For the saved data, the instrument allows you to: • View detailed data for each sample (page 32). • Rename data files (page 35). •...
  • Page 35 On the Export data screen, press the data set of interest. A Data screen displaying a list of data entries for that data set appears. To view additional data entries that do not fit in the screen, scroll up or down. Data screen for quantitation assays Data screen for quality assays To reverse the order of the data entries, press the blue chevron above the...
  • Page 36 To view the sample details, press the sample of interest. A Data details screen opens. Data details for quantitation assays Data details for quality assays Qubit 4 Fluorometer User Guide ™...

  • Page 37: Rename Data Files

    Rename data files Rename data files On the Concentration, Graph, RNA IQ results, or Home screen, press Data. On the Export data screen, select the data set of interest. From the list of data entries for the selected data set, press the sample of interest to open the Data details screen.

  • Page 38: Export Data

    Export data Introduction The Qubit ™ 4 Fluorometer is designed for standalone use; it does not require an external computer. However, to archive data and generate reports, you can export the numeric data stored in the CSV file to a computer either by using the USB drive or by saving directly to the computer using the USB cable.
  • Page 39 Press Export to export the data. The numeric data is automatically saved as a CSV file.  Transfer the USB drive to the USB drive port on your computer. You can open the CSV file using any spreadsheet program. Export data directly Data files may also be exported from the Qubit ™...
  • Page 40 Information saved to the CSV file Note: For the RNA IQ assay, there is no original sample concentration, Qubit ™ tube concentration or dilution factor. Instead, the results file contains % large and % small RNA information. Qubit 4 Fluorometer User Guide ™...

  • Page 41: Delete Data

    Delete data Delete data files On the Concentration, Graph, RNA IQ results, or Home screen, press Data. On the Export data screen, check the selection box to the left of each data set you wish to delete. To delete only individual data entries from a data set, press the data set of interest, and then check the selection box to the left of the samples you wish to delete.

  • Page 42: Manage, Import, And Create Assays

    5. Manage, import, and create assays Overview The Qubit 4 Fluorometer allows you to personalize the instrument’s user ™ interface to hide and show (unhide) assays or folders and to move them to different locations. In addition, you can add custom MyQubit assays by importing them from our website at thermofisher.com/qubit or create new assays using the MyQubit software and web tool.

  • Page 43: Show/Hide Assays

    Show/Hide assays Show/Hide all From the Settings screen, press Manage assays. assays within the On the Manage assays screen, press Show/hide assays. assay folder The software displays a list of the existing assay folders available on the instrument. Each assay folder contains individual assays. ...
  • Page 44 Show/Hide From the Settings screen, press Manage assays. individual assays On the Manage assays screen, press Show/hide assays. The software displays a list of the existing assay folders available on the instrument. Each assay folder contains individual assays.  Select the assay folder containing the assays you wish to show or hide. The software displays a list of existing individual assays within the selected assay folder and the controls for showing and hiding them.

  • Page 45: Move Assays

    Move assays Move assay folders/ From the Settings screen, press Manage assays. individual assays on On the Manage assays screen, press Move assays. the Home screen The software displays the Manage assays screen with available assay folders and individual assays, and activates the move function. ...
  • Page 46 Move assays From the Settings screen, press Manage assays. between folders On the Manage assays screen, press Move assays. The software displays the Manage assays screen with available assay folders and individual assays, and activates the move function.  Press the desired assay folder to open the folder’s assay screen. Press and hold the individual assay to detach it and drag it over the Home screen icon ( Release the drag to drop the selected assay to the Home screen.

  • Page 47: Import Assays

    Import assays MyQubit assay files You can add pre-existing MyQubit assays to your Qubit ™ 4 Fluorometer by downloading the assay files from thermofisher.com/qubit, then importing the files to the instrument from a USB drive or from an external computer via a USB cable. Note: MyQubit assay files have the file extension .qbt.
  • Page 48 Press Next. The Import new assay screen displays the name of the assay being imported (or the first assay, if multiple assays were selected) and lists the destination folders where you can save it. Press the Back button ( ) or Cancel to return to the Settings screen without importing the assays.
  • Page 49 In the New folder screen, press the New folder text field. The keyboard opens. Using the keyboard, enter the name of the new destination folder. You can use up to 11 characters; spaces at the beginning or the end of the name are not recognized.
  • Page 50 Select the destination folder for the next assay in your import queue, then press Save. Repeat this process until you have imported all the assays you have selected in Step 4 (page 45). Once all the assays you have selected are imported successfully, the Verification screen opens.
  • Page 51 Import assays from When the Qubit ™ 4 Fluorometer is connected to a computer via a USB cable, MyQubit .qbt assay files can be transferred directly from the computer to the a computer instrument. Go to thermofisher.com/qubit and download the MyQubit assay files (.qbt file) you wish to install.

  • Page 52: Create Your Own Assay For The Qubit

    Create your own assay for the Qubit 4 Fluorometer ™ Create a MyQubit All Qubit ™ 4 Fluorometers are pre-loaded with MyQubit software, which allows you to create new applications for the Qubit ™ 4 Fluorometer using parameters assay uploaded to the instrument via a USB drive, without changing the existing assays. Fluorometer mode allows you to use the Qubit 4 Fluorometer with any reagent ™...

  • Page 53: Instrument Maintenance

    6. Instrument maintenance Maintenance and cleaning Maintenance The Qubit 4 Fluorometer does not need regular maintenance. To troubleshoot ™ problems with the instrument, contact Technical Support (page 69). • Do not perform any repairs or service on the Qubit 4 Fluorometer to avoid ™...

  • Page 54: Software Updates

    Software updates Update the To update the Qubit ™ 4 Fluorometer with the latest software from Thermo Fisher Scientific: software Download the latest software to a USB drive or to your computer from thermofisher.com/qubit. If using a USB drive, insert the USB drive into the instrument. If using a computer, connect the Qubit ™...
  • Page 55 When the appropriate files are detected, the Software update screen is displayed. Press Update to update the software.  When prompted, press Restart to complete the software update. Qubit 4 Fluorometer User Guide ™...

  • Page 56: Device Verification Test

    Device verification test Perform Device The device verification test checks the internal components of the Qubit ™ Fluorometer. Perform the device verification test when a problem with the verification test instrument is suspected. It is not necessary to perform the verification test on a regular basis.

  • Page 57: Appendix A: Troubleshooting

    Appendix A: Troubleshooting Troubleshooting Handling samples • The calibration standards included in the Qubit ™ microRNA, Qubit ™ RNA HS, Qubit RNA XR, Qubit RNA IQ, and Qubit RNA BR Assay Kits are high- ™ ™ ™ quality RNA standards. The integrity and concentration of these standards is critical to the optimal performance of the Qubit RNA assays.
  • Page 58 Low reading • The sample is out of range. Use a sample that is more concentrated or use a lower dilution (for example, 20 µL in 180 µL instead of 10 µL in 190 µL). • For Qubit quantitation assays, view the ™...

  • Page 59: Critical Qubit ™ Assay Considerations

    Critical Qubit Assay considerations ™ How the Qubit The Qubit ™ 4 Fluorometer generates concentration data based on the relationship ™ between the two standards used in calibration (three for the Qubit ™ protein Fluorometer assay). The plot below shows the line corresponding to the curve-fitting calculates algorithm (a modified Hill plot) used in the calculation of concentration data for concentration...
  • Page 60 Assay temperature The Qubit ™ assays were designed to be performed at room temperature (22–28°C), and temperature fluctuations can influence the accuracy of the assay. To minimize temperature fluctuations, store all kit reagents at room temperature and insert all assay tubes into the Qubit 4 Fluorometer only for as much time as ™...

  • Page 61: Appendix B: Ordering Information

    Appendix B: Ordering information Qubit 4 Fluorometer and accessories ™ The following products can be used with the Qubit 4 Fluorometer and are ™ available separately from Thermo Fisher Scientific. For more information, visit thermofisher.com or contact Technical Support (page 69). Product Quantity Cat.

  • Page 62: Appendix C: Safety

    Appendix C: Safety Symbols on instruments Electrical symbols The following table describes the electrical symbols that may be displayed. Symbol Description Indicates a terminal that can receive or supply alternating current or voltage. Safety symbols The following table describes the safety symbols that may be displayed. Each symbol may appear by itself or in combination with text that explains the relevant hazard (see “...

  • Page 63: Safety Labels On Instruments

    Safety labels on instruments The following CAUTION, WARNING, and DANGER statements may be displayed on Thermo Fisher Scientific instruments in combination with the safety symbols described in the preceding section. Hazard English Français symbol CAUTION! Hazardous chemicals. Read the ATTENTION! Produits chimiques Safety Data Sheets (SDSs) before handling.

  • Page 64: General Instrument Safety

    General instrument safety WARNING! PHYSICAL INJURY HAZARD. Use this product only as specified in this document. Using this instrument in a manner not specified may result in personal injury or damage to the instrument. Operating the Ensure that anyone who operates the instrument has: instrument •...

  • Page 65: Chemical Safety

    Chemical safety Chemical hazard WARNING! CHEMICAL HAZARD. Before handling any chemicals, refer warning to the Safety Data Sheet (SDS) provided by the manufacturer, and observe all relevant precautions. WARNING! CHEMICAL STORAGE HAZARD. Never collect or store waste in a glass container because of the risk of breaking or shattering. Reagent and waste bottles can crack and leak.

  • Page 66: Chemical Waste Safety

    Chemical waste safety Chemical waste CAUTION! HAZARDOUS WASTE. Refer to Safety Data Sheets (SDSs) hazard and local regulations for handling and disposal. Chemical waste To minimize the hazards of chemical waste: safety guidelines • Read and understand the Safety Data Sheets (SDSs) provided by the manufacturers of the chemicals in the waste container before you store, handle, or dispose of chemical waste.

  • Page 67: Electrical Safety

    Electrical safety DANGER! ELECTRICAL SHOCK HAZARD. Severe electrical shock can result from operating the Qubit ™ 4 Fluorometer without its instrument panels in place. Do not remove instrument panels. High‐voltage contacts are exposed when instrument panels are removed from the instrument. Power DANGER! ELECTRICAL HAZARD.

  • Page 68: Biological Hazard Safety

    Biological hazard safety WARNING! BIOHAZARD. Biological samples such as tissues, body fluids, and blood of humans and other animals have the potential to transmit infectious diseases. Follow all applicable local, state/provincial, and/or national regulations. Wear appropriate protective eyewear, clothing, and gloves. Read and follow the guidelines in these publications. ATTENTION! BIOHAZARD.

  • Page 69: Safety And Electromagnetic Compatibility (Emc) Standards

    Safety and Electromagnetic Compatibility (EMC) standards This section provides information on: • U.S. and Canadian safety and EMC standards • European safety and EMC standards • Australian EMC standards U.S. and Canadian The Qubit 4 Fluorometer has been tested to and complies with standard: ™...
  • Page 70 Australian EMC This instrument has been tested to and complies with standard AS/NZS CISPR 11, “Limits and Methods Measurement of Electromagnetic Disturbance Characteristics standards of Industrial, Scientific, and Medical (ISM) Radio‐ frequency Equipment.” Qubit 4 Fluorometer User Guide ™...

  • Page 71: Documentation And Support

    Documentation and support Obtaining support Technical support For the latest services and support information for all locations, visit thermofisher.com. At the website, you can: • Access worldwide telephone and fax numbers to contact Technical Support and Sales facilities • Search through frequently asked questions (FAQs) •...
  • Page 72 thermofisher.com/support | thermofisher.com/askaquestion thermofisher.com 05 October 2017...

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